Toothpaste composition for alleviating dentin hyperesthesia

ABSTRACT

A toothpaste composition for alleviating dentin hyperesthesia is disclosed. The composition includes a peptide consisting of an amino acid sequence of the following Formula 1: 
       K-Y-R1-R2-R3-R4-R5-R6-R7-R8  (Formula 1)
         wherein R1 is arginine (R), lysine (K) or glutamine (Q);   R2 is arginine (R) or glutamine (Q);   R3, R4, and R5 are arginine (R) or lysine (K), respectively;   R6 is asparagine (N) or serine (S); and   R7 and R8 are lysine (K) or tyrosine (Y), respectively.

BACKGROUND OF THE INVENTION 1. Field of the Invention

The present invention relates to toothpaste composition, and morespecifically, the present invention relates to toothpaste compositionfor preventing or alleviating dentin hyperesthesia by inducingphysiological remineralization of defects in dentin constituting atooth.

2. Description of the Related Art

Dentin hyperesthesia, commonly referred to as ‘sensitive dentin’, is acommon symptom experienced by 8% to 57% of the adult population. Inparticular, the case of periodontal disease, which is the most commondisease in Korea, 72.5% to 98% of patients suffer from ‘sensitivedentin’ (Source: National Health Information Portal Medical Information;health.cdc.go.kr/health/Main.do).

Dentin hyperesthesia can be defined as a pain caused by the exposure ofdentinal tubules transmitting all external stimuli to the pulp nerve.This makes the pulp nerve to be more sensitive to the same stimulus.Although there are no nerves in the dentin itself, but we can perceivethe changes because the cold temperature stimulus is transmitted throughthe dentinal tubules to the nerves inside the pulp.

In the dentin, which occupies the most part of the tooth, there aredentinal tubules that extend from the pulp to the enamel. These tubulesare filled with liquid. The diameter increases toward the pulp and has adense structure. Because of these distinct structures, external stimulican be transmitted quickly to the pulp nerve. If the dentin surface isdamaged and the number of exposed dentinal tubules increases, it maycause a more sensitive reaction to the same stimulus than usual.

Currently, there are two ways for dentin hyperesthesia, depending on theprinciple of action. One is to interfere with the signal transmission ofthe nerve that transmits pain, and the other is to block the exposeddentinal tubules to alleviate the symptoms.

Dipotassium phosphate (K₂HPO₄) has been widely used in a method forinterfering with the signal transmission of nerves that transmit pain.However, dipotassium phosphate has a low pain-blocking effect and mustbe used repeatedly, and it is not an effective treatment method becauseit limits the sense of chewing.

Next, calcium hydrogen phosphate (CaHPO₄), fluorine, oxalate, arginine(amino acid), and calcium carbonate (CaCO₃) are used to block thedentinal tubules. In recent years, considering the inconvenience ofreceiving treatment at the dentist for dentin hypersensitivity,toothpastes for preventing or alleviating dentin sensitivity are on themarket, including calcium phosphate, dental type silica, strontiumchloride, calcium carbonate or tricalcium phosphate, as activeingredients. However, these are also foreign materials different fromthe original dentin, so there would be a gap created in the peripheralboundary area between dentin and the foreign materials. The nerve wouldbe exposed again after the foreign material being separated from thesealing, and it would cause a recurrence of dentin sensitivity.

SUMMARY OF THE INVENTION

Embodiments of the present inventive concepts may provide toothpastecomposition for alleviating dentin hyperesthesia, comprising a peptideincluding an amino acid sequence of the following Formula 1:

K-Y-R1-R2-R3-R4-R5-R6-R7-R8  (Formula 1)

wherein R1 is arginine (R), lysine (K) or glutamine (Q);

R2 is arginine (R) or glutamine (Q);

R3, R4, and R5 are arginine (R) or lysine (K), respectively;

R6 is asparagine (N) or serine (S); and

R7 and R8 are lysine (K) or tyrosine (Y), respectively.

Embodiments of the present inventive concepts may also provide acomposition for oral care comprising a peptide including any one aminoacid sequence of SEQ ID NOS: 1 to 96.

Embodiments of the present inventive concepts, 0.0015-0.0025 parts byweight of the peptide may be included based on 100 parts by weight ofthe composition.

Embodiments of the present inventive concepts, 31.00-33.00 parts byweight of tricalcium phosphate may be included based on 100 parts byweight of the composition.

Embodiments of the present inventive concepts, 0.045-0.055 parts byweight of hydroxyapatite may be included based on 100 parts by weight ofthe composition.

Embodiments of the present inventive concepts, the toothpastecomposition may include 20-22 parts by weight of purified water,humectant 33-37 parts by weight, viscosity modifier 4.4-5.4 parts byweight, surfactant 1.1-1.3 parts by weight, flavoring agent 0.8-0.9parts by weight, and diluting agent 0.04-0.06 parts by weight based on100 parts by weight of the composition.

Embodiments of the present inventive concepts, the toothpastecomposition may include 0.15-0.25 parts by weight of aminocaproic acidand 1.9-2.1 parts by weight of allantoin based on 100 parts by weight ofthe composition.

Embodiments of the present inventive concepts, the humectant may beD-sorbitol solution or sodium PCA solution, or concentrated glycerin,the viscosity modifier may be hydrous silicic acid, xanthan gum orCMC(Carboxymethyl Cellulose Sodium Salt), the surfactant may be sodiumcocoylmethyltaurate, flavoring agent is green tea extract, chamomileextract, rosemary extract, myrrh tincture, rhatany tincture, chamomiletincture, mastic oil 40 HF-60662, propolis extract, grapefruit seedextract, spearmint B71228 or peppermint oil 81689, and the dilutingagent may be hydroxyapatite.

Embodiments of the present inventive concepts, the humectant may include80-83% by weight of the D-sorbitol solution, 7-8% by weight of theSodium PCA solution, and 10-12% by weight of the concentrated glycerin.

Embodiments of the present inventive concepts, the viscosity modifiermay include 79-85% by weight of the hydrous silicic acid, 4-8% by weightof xanthan gum, and 11-13% by weight of the CMC.

Effect of the Invention

The present inventive concepts may provide toothpaste composition thatprevents or alleviates dentin hyperesthesia by sealing off exposeddefects of the dentinal tubule through physiological remineralization ofdentin.

Other aspects, advantages and salient features of the embodiments willbecome apparent to those skilled in the art from the following detaileddescription, which, taken in conjunction with the annexed drawings,discloses exemplary embodiments of the disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a graph showing the results of comparing the effect of therespective groups of the peptide included in the toothpaste compositionfor alleviating dentin hyperesthesia according to an embodiment of thepresent invention on the expression of DSPP, which is an odontoblastdifferentiation marker gene.

FIG. 1B is a graph showing the results of comparing the expressionlevels of the odontoblast differentiation marker Dspp gene in MDPC-23cells treated with the peptides included in the toothpaste compositionfor alleviating dentin hyperesthesia according to an embodiment of thepresent invention.

FIG. 1C is a graph showing the results of comparing expression levels ofodontoblast differentiation marker genes, Dspp, Dmp1, and Nestin inMDPC-23 cells treated with peptides of Group 11 and Group 12 included inthe toothpaste composition for alleviating dentin hyperesthesiaaccording to an embodiment of the present invention.

FIG. 1D is a graph showing the results of evaluating cytotoxicity of thepeptides included in the toothpaste composition for alleviating dentinhyperesthesia according to an embodiment of the present invention.

FIG. 2 is a result of measuring the molecular weight through MALDI-TOFanalysis to confirm the stability of the peptide contained in thetoothpaste composition for relieving dentin hyperesthesia according toan embodiment of the present invention. In FIG. 2, A shows the molecularweight of the peptide itself contained in the toothpaste composition forrelieving dentin hyperesthesia according to an embodiment of the presentinvention. In FIG. 2, B shows the molecular weight of the peptidecontained in the state of the toothpaste composition for relievingdentin hyperesthesia according to an embodiment of the presentinvention.

FIG. 3 shows the permeability of toothpaste composition for alleviatingdentin hyperesthesia according to an embodiment of the presentinvention. A fluorescence dyeing reagent (Rhodamine B) was mixed andtreated for 1 minute on the tooth exposed to the dentinal tubules andthen observed with a fluorescence microscope.

FIG. 4 shows the results of comparing the sealing ability of thedentinal tubule with the toothpaste composition for alleviating dentinhyperesthesia according to the embodiment of the present invention(example 3), comparative test example 3-1, and comparative test example3-2.

A-D shows the dentinal tubules treated only with purified water(comparative test example 3-1), E-H is showing the dentinal tubulesbrushed using a toothpaste composition without the peptide comparingwith the embodiment of the present invention (comparative test example3-2), and I-L shows the dentinal tubules brushed using a toothpastecomposition for alleviating dentin hyperesthesia according to theembodiment of the present invention (scale bar: A, E, I, 100 μm; B, F,J, 20 μm; C, G, K, 20 μm; D, H, L, 10 μm). In each case, the dentinslices with exposed dentinal tubules were washed 3 times with distilledwater after brushing once a day for 1 minute and then immersed inartificial saliva. After repeating this process for 2 weeks, the abilityto seal off the dentin tubules was observed with a scanning electronmicroscope.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Reference will now be made in detail to embodiments, examples of whichare illustrated in the accompanying drawings, wherein like referencenumerals refer to like elements throughout. In this regard, the presentembodiments may have different forms and should not be construed aslimited to the descriptions set forth herein. Unless otherwise defined,all terms including technical and scientific terms used herein have thesame meaning as commonly understood by one of ordinary skills in the artto which this disclosure belongs. It will be further understood thatterms, such as those defined in commonly used dictionaries, should beinterpreted as having a meaning that is consistent with their meaning inthe context of the relevant art and will not be interpreted in anidealized or overly formal sense unless expressly so defined herein. Inaddition, terms to be described later are defined in consideration ofcontributions in the present disclosures, which may vary according tothe intention of the user or practice.

The disclosure now will be described more fully hereinafter withreference to the accompanying drawings, in which illustrativeembodiments of the disclosure are shown. This disclosure may, however,be embodied in many different forms and should not be construed aslimited to the embodiments set forth herein; rather, these embodimentsare provided so that this disclosure will be thorough and complete andwill fully convey the scope of the disclosure to those skilled in theart. However, as it is presented as an example, the present invention isnot limited thereto and the present invention is defined only by thescope of the claims which will be described later.

It will be understood that the terms “comprises” and/or “comprising,”when used in this specification, specify the presence of statedcomponents, this means that it may contain more components, rather thanexclude other components, unless there is a particularly contraryarticle.

Hereinafter, embodiments of the present invention are described in moredetail.

An odontoblast may refer to a cell that synthesizes and secretesproteins and polysaccharides composing the matrix of the dentin. It is acolumnar cell that is in contact with the predentin (uncalcified dentin)and forms a cell layer at the periphery of the dental pulp. It is adifferentiated cell (becoming a cell derived from the mesenchymalectoderm) involved in calcification of dentin. At the developmentalstage, an odontoblast faces the enamel among the cells of the dentalpapilla, involved in calcification of dentin.

A peptide, included in the toothpaste composition for alleviating dentinhyperesthesia according to an embodiment of the present invention(hereinafter, ‘odontoblast differentiation promoting peptide’), does notexhibit cytotoxicity, but it is possible to increase the expressionlevel of the odontoblast differentiation marker genes DSPP, Dmp1 andNestin. When transplanted in vivo with pulp tissue cells, the pulptissue cells may form a dentin/dentin-like tissue.

Odontoblast differentiation promoting peptide includes mutant peptideshaving a sequence different from the amino acid sequence constitutingthe amino acid sequence and at least one amino acid residue, as long asit can promote dentin regeneration or treat dentin hyperesthesia.

Amino acid exchanges in proteins and polypeptides, which do notgenerally alter the molecular activity, are known in the art. The mostcommonly occurring exchanges are amino acid residues Ala/Ser, Val/Ile,Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Thy/Phe,Ala/Pro, Lys/Arg, Asp/Asn, Leu/Ile, Leu/Val, Ala/Glu, Asp/Gly, in bothdirections. The peptide may include peptides that have improvedstructural stability against heat, pH, etc., or improved ability topromote regeneration of dentin or dental pulp due to alteration ormodification of the amino acid sequence.

For example, although glutamine which is an acidic amino acid atposition 3 of the peptide of SEQ ID NO: 1 of the present invention issubstituted with a basic amino acid, lysine or arginine, the effects ofthe peptide of the present invention may be obtained as it is; althougharginine which is a basic amino acid at position 4 or 5 of the peptideof SEQ ID NO: 1 is substituted with an acidic amino acid glutamine or abasic amino acid lysine, the effects of the peptide of the presentinvention may be obtained as it is; although lysine which is a basicamino acid at position 6, 7, or 9 of the peptide of SEQ ID NO: 1 issubstituted with a basic amino acid arginine or an aromatic amino acidtyrosine, the effects of the peptide of the present invention may beobtained as it is; although asparagine which is an acidic amino acid atposition 8 of the peptide of SEQ ID NO: 1 is substituted with a neutralamino acid serine, the effects of the peptide of the present inventionmay be obtained as it is; and although tyrosine which is an aromaticamino acid at position 10 of the peptide of SEQ ID NO: 1 is substitutedwith a basic amino acid lysine, the effects of the peptide of thepresent invention may be obtained as it is.

As such, although the acidic amino acids, basic amino acids, or aromaticamino acids constituting the peptide of the present invention aresubstituted with amino acids having the same properties, or substitutedwith different acidic amino acids, basic amino acids, neutral aminoacids, or aromatic amino acids, respectively, the effects of the peptideof the present invention may be obtained as it is. Therefore, it isapparent that a peptide variant having a sequence including one or moreamino acid residues different from those of the amino acid sequenceconstituting the peptide of the present invention is also included inthe scope of the peptide of the present invention.

Further, although arbitrary amino acids are added at the N-terminus orC-terminus of the peptide of the prevention, the effects of the peptideof the present invention may be obtained as it is. Therefore, a peptideprepared by adding arbitrary amino acids at the N-terminus or C-terminusof the peptide of the present invention is also included in the scope ofthe peptide of the present invention. For example, a peptide prepared byadding 1 to 300 amino acids at the N-terminus or C-terminus of thepeptide of the present invention may be exemplified, for anotherexample, a peptide prepared by adding 1 to 100 amino acids at theN-terminus or C-terminus of the peptide of the present invention may beexemplified, and for still another example, a peptide prepared by adding1 to 24 amino acids at the N-terminus or C-terminus of the peptide ofthe present invention may be exemplified.

The mRNA of the DSPP gene in MDPC-23 cells treated with the odontoblastdifferentiation promoting peptide, compared to the mRNA level of theDSPP gene in MDPC-23 cells (control) not treated with the odontoblastdifferentiation promoting peptide, was all 1.3 times or more (Tables 13to 24).

As reported up to now, it is known that as the mRNA level of DSPP isincreased, odontoblast differentiation and dentin regeneration arepromoted, and therefore, it can be seen that 128 kinds of peptidesincreases the mRNA level of Dspp gene, which in turn may exhibit theeffect of promoting odontoblast differentiation and dentin regeneration(Taduru Sreenath et al., THE JOURNAL OF BIOLOGICAL CHEMISTRY, Vol. 278,No. 27, Issue of July 4, pp. 24874-24880, 2003; William T. Butler etal., Connective Tissue Research, 44(Suppl. 1): 171-178, 2003).

The peptide included in the toothpaste composition for alleviatingdentin hyperesthesia may be used in a single form of the peptide or apolypeptide form of 2 or more repeats of the peptide, and the peptidemay also be used in a complex form of a drug having a therapeutic effecton dentin or dental pulp diseases linked at the N-terminus or C-terminusof the peptide.

Example 1

Synthesis of Peptides for Promoting Odontoblast Differentiation

The present inventors synthesized a peptide (SEQ ID NO: 1) showing theeffect of promoting regeneration of dentin or dental pulp tissues by a9-fluorenylmethyloxycarbonyl (Fmoc) method, and they synthesizedpeptides of respective groups (Tables 1 to 12) by substituting the aminoacids of the synthesized peptide.

N-KYQRRKKNKY-C(SEQ ID NO: 1)

First, peptides of Group 1 were synthesized by using the peptide of SEQID NO: 1 or by substituting any amino acid at positions 5 to 7 of thepeptide of SEQ ID NO: 1 with lysine or arginine (Table 1).

TABLE 1 Peptides of Group 1 SEQ ID NO: Amino acid sequence (N→C) 1KYQRRKKNKY 2 KYQRRKRNKY 3 KYQRRRKNKY 4 KYQRRRRNKY 5 KYQRKKKNKY 6KYQRKRKNKY 7 KYQRKKRNKY 8 KYQRKRRNKY

Next, peptides of Group 2 were synthesized by substituting any aminoacid at positions 5 to 7 of the peptide of SEQ ID NO: 1 with lysine orarginine or by substituting an amino acid at position 8 of the peptideof SEQ ID NO: 1 with serine (Table 2).

TABLE 2 Peptides of Group 2 SEQ ID NO: Amino acid sequence (N→C) 9KYQRRKKSKY 10 KYQRRKRSKY 11 KYQRRRKSKY 12 KYQRRRRSKY 13 KYQRKKKSKY 14KYQRKRKSKY 15 KYQRKKRSKY 16 KYQRKRRSKY

Next, peptides of Group 3 were synthesized by substituting any aminoacid at positions 5 to 7 of the peptide of SEQ ID NO: 1 with lysine orarginine or by substituting an amino acid at position 9 of the peptideof SEQ ID NO: 1 with tyrosine (Table 3).

TABLE 3 Peptides of Group 3 SEQ ID NO: Amino acid sequence (N→C) 17KYQRRKKNYK 18 KYQRRKRNYK 19 KYQRRRKNYK 20 KYQRRRRNYK 21 KYQRKKKNYK 22KYQRKRKNYK 23 KYQRKKRNYK 24 KYQRKRRNYK

Next, peptides of Group 4 were synthesized by substituting any aminoacid at positions 5 to 7 of the peptide of SEQ ID NO: 1 with lysine orarginine, by substituting an amino acid at position 8 of the peptide ofSEQ ID NO: 1 with serine, by substituting an amino acid at position 9 ofthe peptide of SEQ ID NO: 1 with tyrosine, or by substituting an aminoacid at position 10 of the peptide of SEQ ID NO: 1 with lysine (Table4).

TABLE 4 Peptides of Group 4 SEQ ID NO: Amino acid sequence (N→C) 25KYQRRKKSYK 26 KYQRRKRSYK 27 KYQRRRKSYK 28 KYQRRRRSYK 29 KYQRKKKSYK 30KYQRKRKSYK 31 KYQRKKRSYK 32 KYQRKRRSYK

Next, peptides of Group 5 were synthesized by substituting an amino acidat position 3 of the peptide of SEQ ID NO: 1 with arginine, bysubstituting an amino acid at position 4 of the peptide of SEQ ID NO: 1with glutamine, or by substituting any amino acid at positions 5 to 7 ofthe peptide of SEQ ID NO: 1 with lysine or arginine (Table 5).

TABLE 5 Peptides of Group 5 SEQ ID NO: Amino acid sequence (N→C) 33KYRQRKKNKY 34 KYRQRKRNKY 35 KYRQRRKNKY 36 KYRQRRRNKY 37 KYRQKKKNKY 38KYRQKRKNKY 39 KYRQKKRNKY 40 KYRQKRRNKY

Next, peptides of Group 6 were synthesized by substituting an amino acidat position 3 of the peptide of SEQ ID NO: 1 with arginine, bysubstituting an amino acid at position 4 of the peptide of SEQ ID NO: 1with glutamine, by substituting any amino acid at positions 5 to 7 ofthe peptide of SEQ ID NO: 1 with lysine or arginine, or by substitutingan amino acid at position 8 of the peptide of SEQ ID NO: 1 with serine(Table 6).

TABLE 6 Peptides of Group 6 SEQ ID NO: Amino acid sequence (N→C) 41KYRQRKKSKY 42 KYRQRKRSKY 43 KYRQRRKSKY 44 KYRQRRRSKY 45 KYRQKKKSKY 46KYRQKRKSKY 47 KYRQKKRSKY 48 KYRQKRRSKY

Next, peptides of Group 7 were synthesized by substituting an amino acidat position 3 of the peptide of SEQ ID NO: 1 with arginine, bysubstituting an amino acid at position 4 of the peptide of SEQ ID NO: 1with glutamine, by substituting any amino acid at positions 5 to 7 ofthe peptide of SEQ ID NO: 1 with lysine or arginine, by substituting anamino acid at position 9 of the peptide of SEQ ID NO: 1 with tyrosine,or by substituting an amino acid at position 10 of the peptide of SEQ IDNO: 1 with lysine (Table 7).

TABLE 7 Peptides of Group 7 SEQ ID NO: Amino acid sequence (N→C) 49KYRQRKKNYK 50 KYRQRKRNYK 51 KYRQRRKNYK 52 KYRQRRRNYK 53 KYRQKKKNYK 54KYRQKRKNYK 55 KYRQKKRNYK 56 KYRQKRRNYK

Next, peptides of Group 8 were synthesized by substituting an amino acidat position 3 of the peptide of SEQ ID NO: 1 with arginine, bysubstituting an amino acid at position 4 of the peptide of SEQ ID NO: 1with glutamine, by substituting any amino acid at positions 5 to 7 ofthe peptide of SEQ ID NO: 1 with lysine or arginine, by substituting anamino acid at position 8 of the peptide of SEQ ID NO: 1 with serine, bysubstituting an amino acid at position 9 of the peptide of SEQ ID NO: 1with tyrosine, or by substituting an amino acid at position 10 of thepeptide of SEQ ID NO: 1 with lysine (Table 8).

TABLE 8 Peptides of Group 8 SEQ ID NO: Amino acid sequence (N→C) 57KYRQRKKSYK 58 KYRQRKRSYK 59 KYRQRRKSYK 60 KYRQRRRSYK 61 KYRQKKKSYK 62KYRQKRKSYK 63 KYRQKKRSYK 64 KYRQKRRSYK

Next, peptides of Group 9 were synthesized by substituting an amino acidat position 3 of the peptide of SEQ ID NO: 1 with lysine, bysubstituting an amino acid at position 4 of the peptide of SEQ ID NO: 1with glutamine, or by substituting any amino acid at positions 5 to 7 ofthe peptide of SEQ ID NO: 1 with lysine or arginine (Table 9).

TABLE 9 Peptides of Group 9 SEQ ID NO: Amino acid sequence (N→C) 65KYKQRKKNKY 66 KYKQRKRNKY 67 KYKQRRKNKY 68 KYKQRRRNKY 69 KYKQKKKNKY 70KYKQKRKNKY 71 KYKQKKRNKY 72 KYKQKRRNKY

Next, peptides of Group 10 were synthesized by substituting an aminoacid at position 3 of the peptide of SEQ ID NO: 1 with lysine, bysubstituting an amino acid at position 4 of the peptide of SEQ ID NO: 1with glutamine, by substituting any amino acid at positions 5 to 7 ofthe peptide of SEQ ID NO: 1 with lysine or arginine, or by substitutingan amino acid at position 8 of the peptide of SEQ ID NO: 1 with serine(Table 10).

TABLE 10 Peptides of Group 10 SEQ ID NO: Amino acid sequence (N→C) 73KYKQRKKSKY 74 KYKQRKRSKY 75 KYKQRRKSKY 76 KYKQRRRSKY 77 KYKQKKKSKY 78KYKQKRKSKY 79 KYKQKKRSKY 80 KYKQKRRSKY

Next, peptides of Group 11 were synthesized by substituting an aminoacid at position 3 of the peptide of SEQ ID NO: 1 with lysine, bysubstituting an amino acid at position 4 of the peptide of SEQ ID NO: 1with glutamine, by substituting any amino acid at positions 5 to 7 ofthe peptide of SEQ ID NO: 1 with lysine or arginine, by substituting anamino acid at position 9 of the peptide of SEQ ID NO: 1 with tyrosine,or by substituting an amino acid at position 10 of the peptide of SEQ IDNO: 1 with lysine (Table 11).

TABLE 11 Peptides of Group 11 SEQ ID NO: Amino acid sequence (N→C) 81KYKQRKKNYK 82 KYKQRKRNYK 83 KYKQRRKNYK 84 KYKQRRRNYK 85 KYKQKKKNYK 86KYKQKRKNYK 87 KYKQKKRNYK 88 KYKQKRRNYK

Lastly, peptides of Group 12 were synthesized by substituting an aminoacid at position 3 of the peptide of SEQ ID NO: 1 with lysine, bysubstituting an amino acid at position 4 of the peptide of SEQ ID NO: 1with glutamine, by substituting any amino acid at positions 5 to 7 ofthe peptide of SEQ ID NO: 1 with lysine or arginine, by substituting anamino acid at position 8 of the peptide of SEQ ID NO: 1 with serine, bysubstituting an amino acid at position 9 of the peptide of SEQ ID NO: 1with tyrosine, or by substituting an amino acid at position 10 of thepeptide of SEQ ID NO: 1 with lysine (Table 12).

TABLE 12 Peptides of Group 12 SEQ ID NO: Amino acid sequence (N→C) 89KYKQRKKSYK 90 KYKQRKRSYK 91 KYKQRRKSYK 92 KYKQRRRSYK 93 KYKQKKKSYK 94KYKQKRKSYK 95 KYKQKKRSYK 96 KYKQKRRSYK

Example 2: Verification of the Effect of Promoting Regeneration ofDentin Using the Odontoblasts Example 2-1: Validation of the Effect ofPeptides on the Activity of the DSPP (Dentin Sialophosphoprotein)Promoter

First, MDPC-23 cells, which are mouse-derived odontoblasts, werecultured in DMEM medium containing 10% FBS, 5% CO2 and 37° C.

Next, the cultured MDPC-23 cells were dispensed into a 24-well plate at5×10⁴ cells per well, incubated for 24 hours. And then usingLipofectamine Plus™ reagent, the cultured cells were transformed byintroducing a recombinant (pGL3 vector) the DSPP promoter and luciferasegene were introduced. The transformed MDPC-23 cells were treated withthe peptides of groups 1 to 12 synthesized in Example 1, respectively,and cultured for 48 hours. Then luciferase activity was measured, andthe average level was compared (FIG. 1A). As a control, transformedMDPC-23 cells without an odontoblast differentiation promoting peptidewere used.

FIG. 1A is a graph showing the effect of each peptide provided in thepresent invention on the expression of DSPP, an odontoblastdifferentiation marker gene, for each group. As shown in FIG. 1A, eachpeptide provided in the present invention showed a value of about 1.3times or more of the luciferase activity level measured in the controlgroup as a whole, but the difference was shown for each group, and thepeptide of group 12 was the highest. And the next highest level ofluciferase activity was from group 11 peptide. Therefore, it wasverified that the peptides provided by the present invention exhibit aneffect of activating the DSPP promoter.

Example 2-2: Verification of the Effect of Peptides on the ExpressionLevel of the DSPP Gene, an Odontoblast Differentiation Marker Gene

The MDPC-23 cells cultured in Example 2-1 were treated with the peptidesof each group synthesized in Example 1, then cultured for 48 hours. ThemRNA level of the DSPP, an odontoblast differentiation marker gene,expressed in the MDPC-23 cells were measured, and the measured mRNAlevel of each DSPP gene was converted into a relative ratio to the mRNAlevel of the DSPP gene measured in control (Tables 13 to 24). Inaddition, the average value of the mRNA level of the DSPP gene measuredaccording to the peptides of each group was compared (FIG. 1B). At thistime, as a control, MDPC-23 cells that were not treated with the peptidepromoting differentiation of odontoblast were used.

The expression level of the DSPP gene was measured through RT-PCR andreal-time PCR analysis: Specifically, total RNA was extracted from theMDPC-23 cells using TRIzol reagent. 2 μg of the total RNA, 1 μl ofreverse transcriptase, and 0.5 μg of oligo (oligo; dT) were used tosynthesize cDNA. The synthesized cDNA was used in a real-time polymerasechain reaction. The real-time polymerase chain reaction was performed onan ABI PRISM 7500 sequence detection system (Applied Biosystems) and anSYBR GREEN PCR Master Mix (Takara, Japan). The real-time polymerasechain reaction was performed under conditions of 94° C., 1 min; 95° C.,15 sec; 60° C., 34 sec for 40 cycles. Results were analyzed by acomparative cycle threshold (CT) method. At this time, the Gapdh genewas used as the internal control group, and the measured value wasrepeated three times. The mean value and standard deviation valuethereof were used.

Dspp_F: (SEQ ID NO: 97) 5'-ATTCCGGTTCCCCAGTTAGTA-3' Dspp_R:(SEQ ID NO: 98) 5'-CTGTTGCTAGTGGTGCTGTT-3' Gapdh_F: (SEQ ID NO: 99)5'-AGGTCGGTGTGAACGGATTTG-3' Gapdh_R: (SEQ ID NO: 100)5'-TGTAGACCATGTAGTTGAGGTCA-3'

TABLE 13 Effects of peptides of group 1 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 1 1.754 0.132 21.646 0.092 3 1.464 0.221 4 1.855 0.102 5 1.639 0.057 6 1.746 0.091 71.864 0.132 8 1.639 0.032

TABLE 14 Effects of peptides of group 2 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 9 1.854 0.032 101.746 0.052 11 1.639 0.201 12 1.548 0.027 13 1.685 0.077 14 1.846 0.14115 1.964 0.279 16 1.739 0.092

TABLE 15 Effects of peptides of group 3 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 17 2.117 0.209 182.319 0.092 19 1.931 0.102 20 2.553 0.099 21 1.893 0.132 22 2.412 0.07223 2.171 0.281 24 2.212 0.111

TABLE 16 Effects of peptides of group 4 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 25 2.371 0.089 262.193 0.052 27 1.993 0.202 28 2.453 0.192 29 1.883 0.101 30 2.512 0.20931 2.371 0.139 32 2.219 0.302

TABLE 17 Effects of peptides of group 5 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 33 1.712 0.091 341.931 0.172 35 1.983 0.102 36 2.319 0.292 37 1.597 0.301 38 2.116 0.21139 1.712 0.191 40 2.219 0.212

TABLE 18 Effects of peptides of group 6 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 41 1.546 0.091 421.586 0.103 43 1.669 0.095 44 1.793 0.203 45 1.532 0.31 46 1.887 0.07747 1.697 0.009 48 1.558 0.201

TABLE 19 Effects of peptides of group 7 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 49 1.923 0.192 501.887 0.007 51 1.601 0.082 52 2.019 0.135 53 1.592 0.222 54 1.437 0.34155 1.663 0.094 56 1.701 0.109

TABLE 20 Effects of peptides of group 8 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 57 2.039 0.082 581.998 0.172 59 1.792 0.007 60 2.107 0.201 61 2.301 0.019 62 1.672 0.30863 1.769 0.085 64 1.967 0.039

TABLE 21 Effects of peptides of group 9 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 65 1.723 0.072 661.627 0.291 67 1.777 0.027 68 1.432 0.41 69 2.011 0.081 70 1.927 0.10571 1.879 0.06 72 2.011 0.009

TABLE 22 Effects of peptides of group 10 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 73 2.035 0.021 742.011 0.063 75 1.997 0.059 76 2.351 0.109 77 1.729 0.111 78 2.635 0.09179 2.231 0.077 80 1.837 0.201

TABLE 23 Effects of peptides of group 11 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 81 3.092 0.152 823.361 0.098 83 3.572 0.209 84 3.702 0.301 85 3.67 0.088 86 3.705 0.13787 3.888 0.072 88 4.021 0.301

TABLE 24 Effects of peptides of group 12 on mRNA level of Dspp gene mRNAlevel of Dspp gene SEQ ID NO: Mean Standard deviation 89 4.211 0.413 904.811 0.302 91 4.362 0.182 92 4.211 0.287 93 4.525 0.25 94 3.836 0.09995 4.62 0.401 96 5.211 0.371

As shown in Tables 13 to 24, compared to the mRNA level of the DSPP genemeasured in the control group, it was confirmed that the mRNA levels ofthe DSPP gene of the experimental group treated with the peptide wereall 1.3 times or more. In particular, it was confirmed that all thepeptides of group 11 showed a value of 3 times or more in the mRNA levelof the DSPP gene, and all peptides of group 12 showed a value of 3.8times or more in the mRNA level of the DSPP gene.

In addition, FIG. 1B is a graph comparing the expression level of theDSPP gene, an odontoblast differentiation marker, in MDPC-23 cellstreated with an odontoblast differentiation promoting peptide. As shownin FIG. 1B, when the peptide for promoting differentiation ofodontoblast was treated, the mRNA level of the DSPP gene, which is amarker for odontoblast differentiation, is increased. Similar to that ofFIG. 1A, it was confirmed that a value of about 1.3 times or morecompared to the level of DSPP gene mRNA was measured in the controlgroup.

Example 2-3: Verification of the Effect of the Peptide on the ExpressionLevel of the Odontoblast Differentiation Marker Genes DSPP, Dmp1 andNestin Genes

From the results of Example 2-2, it was confirmed that the odontoblastdifferentiation promoting peptide could increase the mRNA level of theDSPP gene, and in particular, the peptides of groups 11 and 12 canincrease the mRNA level of the DSPP gene by at least 3 times or more.

Accordingly, it was confirmed whether the peptides of groups 11 and 12can also increase the mRNA levels of the Dmp1 and Nestin genes, whichare other odontoblast differentiation marker genes.

The following primers were used with the method from Example 2-2. Thepeptides of groups 11 and 12 were used, thereby affecting the expressionlevels of Dmp1 and Nestin genes. The effect of the differentiationpromoting peptide was measured, and the average level was compared (FIG.1C). At this time, as a control group, MDPC-23 cells without a peptidepromoting differentiation of odontoblast were used.

Dmp1_F: (SEQ ID NO 101) 5'-CATTCTCCTTGTGTTCCTTTGGG-3' Dmp1_R:(SEQ ID NO 102) 5'-TGTGGTCACTATTTGCCTGTG-3' Nestin_F: (SEQ ID NO 103)5'-CCCTGAAGTCGAGGAGCTG-3' Nestin_R: (SEQ ID NO 104)5'-CTGCTGCACCTCTAAGCGA-3'

FIG. 1C is a graph showing the results of comparing the expressionlevels of the odontoblast differentiation marker DSPP, Dmp1, and Nestingenes in MDPC-23 cells treated with the peptides of groups 11 and 12. Asshown in FIG. 1c , when the odontoblast differentiation promotingpeptide was treated, the expression levels of the odontoblastdifferentiation marker DSPP, Dmp1, and Nestin genes were all increased,but the level of increase for each gene was different. It was confirmedthat the peptide of group 12 was more effective.

Since each differentiation marker gene is known to be involved in thedifferentiation of odontoblasts and dentin calcification, the peptidesprovided in the present invention were analyzed whether they promote thedentin regeneration.

Example 2-4: Evaluation of Cytotoxicity of Peptides on Pulp Tissue Cells

Human dental pulp cells were separated from wisdom teeth of 10 adults(aged 18-22) at the School of Dentistry, Seoul National University. Indetail, all experiments were performed after the approval of theInstitutional Review Board and obtaining the informed consent frompatients. Wisdom teeth were fractured according to a method of Jung H Set al. (J Mol Histol. (2011)) to expose the dental pulps, and dentalpulp tissues were separated with forceps. Each of the separated dentalpulp tissues was chopped into small pieces with a razor blade, put in a60-mm dish, covered with a coverslip, and then cultured in a Dulbecco'smodified Eagle's medium.

Next, the obtained dental pulp tissue cells were dispensed into a96-well plate so the number of cells per well was to be about 3×10³,cultured for 24 hours. Then the peptides of groups 11 or 12 were treatedat a concentration of 10 or 50 μg/ml. And it was incubated again for 1,3 or 5 days. The cultured cells were washed with PBS, 20 μl of MTTsolution was added, and then reacted at about 37° C. for 4 hours. Afterthe reaction was completed, the MTT solution was removed, 100 μl of DMSOwas added, and absorbance was measured at a wavelength of 540 nm (FIG.1D). At this time, as a control, pulp tissue cells cultured without thepeptide were used.

FIG. 1D is a graph showing the cytotoxicity of a peptide that promotesthe differentiation of oblasts to dental pulp tissue cells. As shown inFIG. 1D, it was confirmed that the survival rate of pulp tissue cellswas at the same level as that of the control group even when theodontoblast differentiation promoting peptide was added.

Example 3: Preparation of Toothpaste Composition for Alleviating DentinHyperesthesia

Step 1

Mixing purified water and D-sorbitol solution

Step 2

Tricalcium phosphate, aminocaproic acid, allantoin, hydrous silicicacid, sodium PCA solution, hydroxyapatite, peptide (SEQ ID NO: 96),enzyme-treated stevia, and xylitol were added and stirred in a stirrerfor about 40 minutes (stirring conditions: PADDLE 10-30 rpm, DISPERSE500-600 rpm, HOMO 2400-3200 rpm).

Step 3

Add (concentrated) glycerin and xanthan gum and stir for about 40minutes (Stirring condition: PADDLE 10-30 rpm, DISPERSE 500-600 rpm,HOMO 2400-3200 rpm).

Step 4:

(concentrated) Glycerin, Carboxymethyl Cellulose Sodium Salt (CMC) addedand stirred for about 40 minutes (Stirring condition: PADDLE 10-30 rpm,DISPERSE 500-600 rpm, HOMO 2400-3200 rpm).

Step 5

Add sodium cocoylmethyltaurate and stirring for about 20 minutes(Stirring conditions: PADDLE 10-30 rpm, DISPERSE 450-650 rpm).

Step 6

Add green tea extract, chamomile extract, rosemary extract, myrrhtincture, rhatany tincture, chamomile tincture, mastic oil 40, propolisextract, grapefruit seed extract, spearmint, peppermint oil, and stirfor about 15 minutes (stirring condition: PADDLE 10-30 rpm, DISPERSE450-650 rpm)

In each step, stirring was carried out under reduced pressure conditions(−760 mmHg).

TABLE 25 Toothpaste composition for alleviating dentin hyperesthesiaaccording to Example 3 Component Ingredient Content (Wt %) 1 SolventPurified water 21.587 humectant D-Sorbitol Solution 30 Staple Tricalciumphosphate 32 Aminocaproic acid 0.2 Allantoin 2 viscosity modifierhydrous silicic acid 4 humectant Sodium PCA solution 3 diluting agentHydroxyapatite 0.05 Odontoblast Peptide (SEQ ID NO: 96) 0.002differentiation promoting peptide sweetening agent enzyme-treated stevia0.1 Xylitol 0.1 2 humectant (concentrated) glycerin 2 viscosity modifierXanthan gum 0.3 3 humectant (concentrated) glycerin 2 viscosity modifierCMC(Carboxymethyl 0.6 Cellulose Sodium Salt) 4 Surfactantcocoylmethyltaurate 1.2 5 Flavoring agent green tea extract 0.01chamomile extract 0.01 rosemary extract 0.01 myrrh tincture 0.01 rhatanytincture 0.01 chamomile tincture 0.01 mastic oil 40 HF-60662 0.001propolis extract 0.05 grapefruit seed extract 0.1 spearmint B71228 0.05Peppermint oil 81689 0.6 Total 100

Preparing Compositions of Comparative Example

Comparative Example 3-1

Prepared purified water of the same volume as in Example 3.

Comparative Example 3-2

Among the ingredients of Example 3, all ingredients other than thosethat did not contain an odontoblast differentiation promoting peptide(SEQ ID NO: 96) were prepared to be contained the same.

Test Example 1

MALDI-TOF analysis of toothpaste composition for alleviating dentinhyperesthesia according to Example 3

A. Preparation of a toothpaste composition solution by dissolving about0.5 g of a sample of the toothpaste composition provided in Example 3 inabout 1 ml of distilled water

B. Centrifuge the solution of A. at about 15,000 rpm for about 10minutes and collect the supernatant

C. 2 ul of the supernatant collected in B. was mixed with 2 ul of thesubstrate solution (10 mg/mL of α-Cyano-4-hydroxycinnamic acid (CHCA) in0.1% TFA/ACN (1:1, v/v)) and MALDI-TOF analysis was performed(ULTRAFLEXIII™ TOF/TOF (Bruker Daltonics)) (FIG. 2, B).

Test Example 2

Observation of the dentinal tubule permeability of the toothpastecomposition for alleviating dentin hyperesthesia according to Example 3

A. Cut the Tooth to Expose the Dentinal Tubules

Cut the crown of the extracted person's tooth horizontally with adiamond saw to expose the dentinal tubules, and then wash twice forabout 5 minutes with a phosphate buffer solution

B. Cleaning Amputated Tooth

The previously cut tooth was reacted with 0.5 Methylenediaminetetraacetic acid (EDTA, pH 7.4) solution for about 5minutes and then washed twice for about 5 minutes with a phosphatebuffer solution.

C. Addition of a Fluorescent Dyeing Reagent to the ToothpasteComposition for Alleviating Dentin Hyperesthesia According to Example 3

Added 0.1% of the fluorescent dyeing reagent to the toothpastecomposition for alleviating dentin hyperesthesia containing theodontoblast differentiation promoting peptide (SEQ ID NO: 96), mixedwell, and then brushed the cut tooth exposed to the dentinal tubules forabout 1 minute.

D. Observation of Penetration of Toothpaste Composition for AlleviatingDentin Hyperesthesia

The brushed cut tooth was washed 3 times with distilled water and thencut lengthwise to a thickness of about 0.5 mm so that the dentinaltubules of the cut tooth looked long using a diamond saw, and the degreeof penetration was observed with a fluorescence microscope (FIG. 3)

Test Example 3

Observation of the sealing ability of the dentinal tubules of thetoothpaste composition for alleviating dentin hyperesthesia according toExample 3

A. Preparation of Artificial Saliva

The composition of artificial saliva is shown in Table 26 below.

The purified water was added to the final concentration of eachcomponent in Table 2 and mixed, and potassium phosphate (K₂HPO₄) wasadded last.

The pH of artificial saliva is measured near 7.2, similar to humansaliva.

TABLE 26 Ingredient concentration (mM) CaC1₂ 0.7 Mgcl₂ 0.2 K₂HPO₄ 4 KCl30 NaN₃ 0.3 HEPES 20

B. Making Dentinal Tubule Specimens

The extracted human tooth was cut horizontally using a diamond saw tomake a 1 mm thick, dentin specimen with exposed dentinal tubules.

The dentin specimen was reacted for about 5 minutes in a 32% phosphoricacid solution to expose the dentinal tubules completely, and then washedthree times with purified water for about 5 minutes. Then, the dentinspecimen was washed 6 times in an ultrasonic cleaner for about 5 minutesto expose the dentinal tubules completely.

Thereafter, washed three times with a phosphate buffer solution andstored.

C. Observation of the Sealing Ability of the Dentinal Tubules of theToothpaste Composition for Alleviating Dentin Hyperesthesia

Using the toothpaste composition for alleviating dentin hyperesthesiaaccording to Example 3, the specimen was brushed for about 1 minute tothe dentinal tubule specimen, washed 3 times with distilled water, andthen reacted to the artificial saliva for about 24 hours.

After repeating this process for 2 weeks, washed three times withdistilled water, dried, and observed the degree of dentinal tubuleblockade with a scanning electron microscope (S-4700™, HITACHI, Tokyo,Japan) (FIG. 4, I-L).

Comparative Test Example 3-1

Using the purified water prepared in Comparative Example 3-1, brushedthe dentinal tubule specimen for about 1 minute, washed 3 times withdistilled water, and then reacted for about 24 hours in artificialsaliva.

After repeating this process for 2 weeks, the specimens were washed withdistilled water 3 times, dried, and observed the degree of dentinaltubule blockade with a scanning electron microscope (FIG. 4, A-D).

Comparative Test Example 3-2

Using the toothpaste prepared in Comparative Example 3-1, brushed thedentinal tubule specimen for about 1 minute, washed 3 times withdistilled water, and then reacted for about 24 hours in artificialsaliva.

After repeating this process for 2 weeks, the specimens were washed withdistilled water 3 times, dried, and observed the degree of dentinaltubule blockade with a scanning electron microscope (FIG. 4, E-H).

Through Test Example 1, the stability of the peptide contained in thetoothpaste composition for alleviating dentin hyperesthesia according tothe embodiment of the present invention can be confirmed. FIG. 2 is aresult of measuring the molecular weight through MALDI-TOF analysis toconfirm the stability of the peptide contained in the toothpastecomposition for alleviating dentin hyperesthesia according to anembodiment of the present invention. In FIG. 2, A shows the molecularweight of the peptide itself contained in the toothpaste composition foralleviating dentin hyperesthesia according to an embodiment of thepresent invention. In FIG. 2, B shows the molecular weight of thepeptide contained in the state of the toothpaste composition foralleviating dentin hyperesthesia according to an embodiment of thepresent invention.

Referring to FIG. 2, it was observed that the peptide contained in thetoothpaste composition according to Example 3 had the same peak(molecular weight of about 1384 Da) as the measured value of themolecular weight, and through this, the stability of mixed dentindifferentiation promoting peptide in the toothpaste composition wasconfirmed (see graph B of FIG. 2).

According to Test Example 2, as a result of observing the dentinaltubule permeability of the toothpaste composition for alleviating dentinhyperesthesia according to Example 3 with a fluorescence microscope, asshown in FIG. 3, in the case of the tooth treated with the composition,fluorescence was strongly observed on the dentin surface. In addition,penetration of the fluorescent staining reagent was observed along thelower side of the exposed dentinal tubules.

Next, the results of comparing Test Example 3 and Comparative TestExamples 3-1 and 3-2 are as shown in FIG. 4. FIG. 4 is a set of imagescomparing the sealing ability of the dentinal tubules of toothpastecomposition for alleviating dentin hyperesthesia according to Example 3,Comparative Example 3-1, and Comparative Example 3-2. And in moredetail, A-D shows the dentinal tubules of the dentin treated only withpurified water (Comparative Example 3-1), and E-H shows the toothpastecomposition without odontoblast differentiation promoting peptide(Comparative Example 3-2), and I-L shows the dentinal tubules reactedwith the composition, including a peptide for oral care that prevents oralleviates dentin hyperesthesia according to an embodiment of thepresent invention. One (size bar: A, E, I, 100 μm; B, F, J, 20 μm; C, G,K, 20 μm; D, H, L, 10 μm).

As can be seen from FIG. 4, it could be observed that the dentinaltubules were blocked by remineralization in the dentinal tubules byreacting with the toothpaste composition for alleviating dentinhyperesthesia according to an embodiment of the present invention.

FIG. 4 is an enlarged image of the dentinal tubules blocked by thecomposition including a peptide for oral care that prevents oralleviates dentin hyperesthesia according to an embodiment of thepresent invention and shows the results of remineralization in theblocked dentinal tubules and dentin surfaces.

As above, the toothpaste composition for alleviating dentinhyperesthesia according to an embodiment of the present invention formsa thin film on the dentin and at the same time strongly binds to thephosphate-calcium ions released from tricalcium phosphate, andremineralizes the exposed dentinal tubules and dentin surfaces. In otherwords, the toothpaste composition for alleviating dentin hyperesthesiaaccording to an embodiment of the present invention inducesremineralization not only on the surface of the exposed dentinal tubulesbut also inside the dentinal tubule, thereby exhibiting the effect ofalleviating and/or preventing dentin hyperesthesia.

While one or more embodiments of the present invention have beendescribed with reference to the figures, it will be understood by thoseof ordinary skill in the art that various changes in form and detailsmay be made therein without departing from the spirit and scope of thepresent invention.

This study was supported by the technology development project of theMinistry of SMEs and Startups in 2017 [S2462696].

1. A toothpaste composition for alleviating dentin hyperesthesia,comprising a peptide consisting of an amino acid sequence of thefollowing Formula 1:K-Y-R1-R2-R3-R4-R5-R6-R7-R8  (Formula 1) wherein R1 is arginine (R),lysine (K) or glutamine (Q); R2 is arginine (R) or glutamine (Q); R3,R4, and R5 are arginine (R) or lysine (K), respectively; R6 isasparagine (N) or serine (S); and R7 and R8 are lysine (K) or tyrosine(Y), respectively, wherein said toothpaste composition includes0.0015-0.0025 parts by weight of the peptide, 31.00-33.00 parts byweight of tricalcium phosphate, 0.045-0.055 parts by weight ofhydroxyapatite, and 20-22 parts by weight of purified water, whereinsaid toothpaste composition forms a thin film on the surface of saiddentin and induces remineralization on said surface of said dentin anddentinal tubule by binding with the phosphate-calcium ions released fromsaid tricalcium phosphate.
 2. The composition of claim 1, wherein saidpeptide is any one amino acid sequence of SEQ ID NOS: 1 to
 96. 3. Thecomposition of claim 1, wherein said toothpaste composition comprises ahumectant 33-37 parts by weight, a viscosity modifier 4.4-5.4 parts byweight, a surfactant 1.1-1.3 parts by weight, and a flavoring agent0.8-0.9 parts by weight, based on 100 parts by weight.
 4. Thecomposition of claim 1, wherein said toothpaste composition comprises0.15-0.25 parts by weight of aminocaproic acid and 1.9-2.1 parts byweight of allantoin, based on 100 parts by weight.
 5. The composition ofclaim 3, wherein said humectant is D-sorbitol solution, sodium PCAsolution, concentrated glycerin, or a mixture thereof, wherein saidviscosity modifier is hydrous silicic acid, xanthan gum, CMC(Carboxymethyl Cellulose Sodium Salt) or a mixture thereof, saidsurfactant is sodium cocoylmethyltaurate, said flavoring agent is greentea extract, chamomile extract, rosemary extract, myrrh tincture,rhatany tincture, chamomile tincture, mastic oil 40 HF-60662, propolisextract, grapefruit seed extract, spearmint B71228, peppermint oil81689, or a mixture thereof.
 6. The composition of claim 5, wherein saidhumectant comprises 80-83% by weight of said D-sorbitol solution, 7-8%by weight of said Sodium PCA solution, and 10-12% by weight of saidconcentrated glycerin.
 7. The composition of claim 5, wherein saidviscosity modifier comprises 79-85% by weight of said hydrous silicicacid, 4-8% by weight of said xanthan gum, and 11-13% by weight of saidCMC.
 8. A method of alleviating dentin hyperesthesia in a subject inneed thereof, administering the toothpaste composition according toclaim 1, to dentin surface of said subject.